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Method for continuously catalyzing and synthesizing ATS-7 by immobilized enzyme fluidization bed

An immobilized enzyme fluidized bed, ATS-7 technology, applied in the field of medicine and chemical industry, can solve the problems of low yield and low utilization rate of enzymes, and achieve the effect of simplifying purification steps, facilitating industrialization, and facilitating separation from products

Pending Publication Date: 2019-06-21
JIANGSU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the defects of low enzyme utilization and low yield in the prior art, the present invention provides a method for continuously catalytically synthesizing ATS-7 in an immobilized enzyme fluidized bed, which not only saves the tedious purification steps , improves the purity of the product, also improves the utilization rate of the enzyme, reduces the production cost, and is suitable for industrial production

Method used

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  • Method for continuously catalyzing and synthesizing ATS-7 by immobilized enzyme fluidization bed
  • Method for continuously catalyzing and synthesizing ATS-7 by immobilized enzyme fluidization bed
  • Method for continuously catalyzing and synthesizing ATS-7 by immobilized enzyme fluidization bed

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Embodiment 1

[0028] A method for continuously catalytically synthesizing ATS-7 in an immobilized enzyme fluidized bed, comprising the steps of:

[0029] 1) Prepare 0.2 mol / L phosphate buffer solution (pH=6.5-7.8) with dipotassium hydrogen phosphate and potassium dihydrogen phosphate, and set aside.

[0030] 2) Add 15g of macroporous silica gel to 35mL, 0.2mol / L phosphate buffer solution and soak for 12h; after soaking, wash the macroporous silica gel: remove the supernatant, add 35mL, 0.2mol / L The fresh phosphate buffer solution, shaking for 3 minutes, standing for 2 minutes, replacing the buffer solution, and repeating the steps of washing the macroporous silica gel 4 times.

[0031] 3) Place the treated macroporous silica gel in 8.5g GDH solution (1.7g GDH is diluted with phosphate buffer solution of 0.2mol / L, pH=7.0, the mass concentration of GDH solution is 20%), and at 25°C, Stir and adsorb at a speed of 200r / min for 55min to obtain immobilized GDH, and the fixation rate of GDH is 60...

Embodiment 2

[0039] A method for continuously catalytically synthesizing ATS-7 in an immobilized enzyme fluidized bed, comprising the steps of:

[0040] 1) Prepare 0.2 mol / L phosphate buffer solution (pH=6.5-7.8) with dipotassium hydrogen phosphate and potassium dihydrogen phosphate, and set aside.

[0041] 2) Add 15g of macroporous silica gel to 35mL, 0.2mol / L phosphate buffer solution and soak for 12h; after soaking, wash the macroporous silica gel: remove the supernatant, add 35mL, 0.2mol / L The fresh phosphate buffer solution, shaking for 3 minutes, standing for 2 minutes, replacing the buffer solution, and repeating the steps of washing the macroporous silica gel 4 times.

[0042] 3) Place the treated macroporous silica gel in 8.5g GDH solution (1.7g GDH is diluted with 0.2mol / L, pH=7.0 phosphate buffer solution, the mass concentration of GDH solution is 16.7%), and at 25°C, Stir and adsorb at a speed of 200r / min for 55min to obtain immobilized GDH, and the fixation rate of GDH is 60%...

Embodiment 3

[0050] A method for continuously catalytically synthesizing ATS-7 in an immobilized enzyme fluidized bed, comprising the steps of:

[0051] 1) Prepare 0.2 mol / L phosphate buffer solution (pH=6.5-7.8) with dipotassium hydrogen phosphate and potassium dihydrogen phosphate, and set aside.

[0052] 2) Add 15g of macroporous silica gel into 35mL, 0.2mol / L phosphate buffer solution and soak for 12h; after soaking, wash the macroporous silica gel: remove the supernatant, add 15g of soaked macroporous silica gel / L of fresh phosphate buffer solution, shake for 3 minutes, let stand for 2 minutes, replace the buffer, and repeat the steps of washing the silica gel 4 times.

[0053] 3) Place the treated macroporous silica gel in 8.5g GDH solution (1.7g GDH is diluted with 0.2mol / L phosphate buffer solution, the mass concentration of GDH solution is 25%), and at 25°C, at 200r / min The mixture was stirred and adsorbed at a rotating speed for 55 minutes to obtain immobilized GDH, and the imm...

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Abstract

The invention belongs to the field of pharmaceutical and chemical industry, and particularly relates to a method for continuously catalyzing and synthesizing ATS-7 by an immobilized enzyme fluidization bed. Mixed liquid of glucose and a nicotinamide adenine dinucleotide phosphate (NADP+) generates reduced nicotinamide adenine dinucleotide phosphate (NADPH) through a GDH (glutamate dehydrogenase) bed, and mixed solution of ATS-6 and generated NADPH is implemented through a CR fluidized bed, so that the ATS-7 is continuously catalyzed and synthesized by the immobilized enzyme fluidization bed. By the continuous reaction catalysis method of the immobilized enzyme fluidization bed, enzyme catalyzing efficiency is improved; enzyme utilization rate is increased, the catalytic efficiency of the immobilized enzyme is improved by 3.1-4.2 times as compared with that of traditional free enzyme, reaction processes can be controlled by independently adjusting different sample feeding flow speed, the immobilized enzyme cannot easily fall off, continuously catalyzing reaction can be achieved, the immobilized enzyme and a product are conveniently separated, the yield of the product is improved, and purification steps are simplified, and industrialization is more facilitated.

Description

technical field [0001] The invention belongs to the field of medicine and chemical industry, and in particular relates to a method for continuously catalytically synthesizing ATS-7 in an immobilized enzyme fluidized bed. Background technique [0002] Atorvastatin (Atorvastatin) was developed by the American company Warner-Lamber (Warner-Lamber) and was approved for marketing in 1997. It is the drug of choice for patients and the first drug with a global sales volume exceeding US$10 billion. [0003] According to the reported synthetic steps of atorvastatin, the most important part is the stereo configuration control of the two hydroxyl groups, so the key intermediate of atorvastatin, 6-cyano-(3R,5R)-di The synthesis of tert-butyl hydroxycaproate (ATS-7) has received extensive attention in recent years. Disclosed in the U.S. Patent (US6867306B2) a kind of chemical synthesis method of target product, this method is raw material with tert-butyl 6-cyano-5-hydroxyl-3-oxoisobuty...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00
Inventor 梁国斌林伟许光斗庄苍伟胡秀秀周文鳞赵文杰
Owner JIANGSU UNIV OF TECH
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